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Expansion Microscopy for Beginners: Visualizing Microtubules in Expanded Cultured HeLa Cells
Chi Zhang1,2, Jeong Seuk Kang3, Shoh M Asano1,2,4
1Media Lab, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts.
Current Protocols in Neuroscience
|June 5, 2020
Summary
Expansion microscopy (ExM) physically expands cells for nanoscale imaging. This protocol details ExM for beginners using HeLa cells and immunostained microtubules, simplifying the technique for new researchers.
Area of Science:
- Biotechnology
- Microscopy
- Cell Biology
Background:
- Expansion microscopy (ExM) enables nanoscale imaging with conventional microscopes.
- ExM physically expands specimens, preserving biomolecular positions.
- Previous ExM protocols exist for proteins and RNAs.
Purpose of the Study:
- To provide a detailed, cookbook-style protocol for Expansion Microscopy.
- To guide beginners and students in performing ExM experiments.
- To facilitate familiarization with ExM experimental setups and required skills.
Main Methods:
- Culturing HeLa cells.
- Immunostaining of microtubules.
- Performing Expansion Microscopy protocol.
- Microscope imaging of expanded cells.
Main Results:
- Successful expansion of cultured HeLa cells.
- Preservation of cellular structure and immunostained microtubules.
- Demonstration of key ExM steps for beginners.
Conclusions:
- The provided ExM protocol is suitable for newcomers and educational settings.
- This method simplifies learning essential ExM techniques.
- ExM offers a pathway to nanoscale resolution imaging for researchers.
